Rabbit Recombinant Monoclonal CDK2 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 239 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IP | WB | ICC/IF | Flow Cyt (Intra) | IHC-P | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested |
Mouse | Expected | Tested | Expected | Expected | Expected |
Rat | Expected | Tested | Expected | Expected | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/40 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 - 1/10000 | Notes - |
Species Rat | Dilution info 1/1000 - 1/10000 | Notes - |
Species Human | Dilution info 1/1000 - 1/10000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/200 | Notes For unpurified use at 1/100. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/80 | Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/50 | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
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Serine/threonine-protein kinase involved in the control of the cell cycle; essential for meiosis, but dispensable for mitosis. Phosphorylates CTNNB1, USP37, p53/TP53, NPM1, CDK7, RB1, BRCA2, MYC, NPAT, EZH2. Triggers duplication of centrosomes and DNA. Acts at the G1-S transition to promote the E2F transcriptional program and the initiation of DNA synthesis, and modulates G2 progression; controls the timing of entry into mitosis/meiosis by controlling the subsequent activation of cyclin B/CDK1 by phosphorylation, and coordinates the activation of cyclin B/CDK1 at the centrosome and in the nucleus. Crucial role in orchestrating a fine balance between cellular proliferation, cell death, and DNA repair in human embryonic stem cells (hESCs). Activity of CDK2 is maximal during S phase and G2; activated by interaction with cyclin E during the early stages of DNA synthesis to permit G1-S transition, and subsequently activated by cyclin A2 (cyclin A1 in germ cells) during the late stages of DNA replication to drive the transition from S phase to mitosis, the G2 phase. EZH2 phosphorylation promotes H3K27me3 maintenance and epigenetic gene silencing. Phosphorylates CABLES1 (By similarity). Cyclin E/CDK2 prevents oxidative stress-mediated Ras-induced senescence by phosphorylating MYC. Involved in G1-S phase DNA damage checkpoint that prevents cells with damaged DNA from initiating mitosis; regulates homologous recombination-dependent repair by phosphorylating BRCA2, this phosphorylation is low in S phase when recombination is active, but increases as cells progress towards mitosis. In response to DNA damage, double-strand break repair by homologous recombination a reduction of CDK2-mediated BRCA2 phosphorylation. Phosphorylation of RB1 disturbs its interaction with E2F1. NPM1 phosphorylation by cyclin E/CDK2 promotes its dissociates from unduplicated centrosomes, thus initiating centrosome duplication. Cyclin E/CDK2-mediated phosphorylation of NPAT at G1-S transition and until prophase stimulates the NPAT-mediated activation of histone gene transcription during S phase. Required for vitamin D-mediated growth inhibition by being itself inactivated. Involved in the nitric oxide- (NO) mediated signaling in a nitrosylation/activation-dependent manner. USP37 is activated by phosphorylation and thus triggers G1-S transition. CTNNB1 phosphorylation regulates insulin internalization. Phosphorylates FOXP3 and negatively regulates its transcriptional activity and protein stability (By similarity). Phosphorylates CDK2AP2 (PubMed:12944431). Phosphorylates ERCC6 which is essential for its chromatin remodeling activity at DNA double-strand breaks (PubMed:29203878).
Cyclin-dependent kinase 2, Cell division protein kinase 2, p33 protein kinase, CDKN2, CDK2
Rabbit Recombinant Monoclonal CDK2 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 239 publications.
Cyclin-dependent kinase 2, Cell division protein kinase 2, p33 protein kinase, CDKN2, CDK2
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
E304
Affinity purification Protein A
The epitope is within the C-terminus of human Cdk2
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 μg)
Lanes 2, 4 and 6: CDK2 knockout HAP1 cell lysate (20 μg)
Lanes 1 and 2: Green signal from target – ab32147 observed at 34 kDa
Lanes 3 and 4: Red signal from loading control – Anti-beta Actin antibody [mAbcam 8226] - Loading Control ab8226 observed at 42 kDa
Lanes 5 and 6: Merged (red and green) signal
ab32147 was shown to specifically react with CDK2 when CDK2 knockout samples were used. Wild-type and CDK2 knockout samples were subjected to SDS-PAGE. ab32147 and Anti-beta Actin antibody [mAbcam 8226] - Loading Control ab8226 (loading control to beta actin) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-Cdk2 antibody [E304] (ab32147)
Predicted band size: 34 kDa
ab32147 (purified) at 1/40 immunoprecipitating Cdk2 from HeLa cells(Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Immunoprecipitation - Anti-Cdk2 antibody [E304] (ab32147)
Predicted band size: 34 kDa
Lanes 1: Wild-type HAP1 cell lysate (20 μg)
Lanes 2: CDK2 knockout HAP1 cell lysate (20 μg)
Lanes 1 - 2: Merged signal (red and green). Green - ab32147 observed at 34 kDa. Red - loading control, Anti-beta Actin antibody [mAbcam 8226] - Loading Control ab8226, observed at 42 kDa or ab18058, observed at 130 kDa.
This western blot image is a comparison between ab32147 and a competitor's top cited rabbit polyclonal antibody.
All lanes: Western blot - Anti-Cdk2 antibody [E304] (ab32147)
Predicted band size: 34 kDa
All lanes: Western blot - Anti-Cdk2 antibody [E304] (ab32147) at 1/5000 dilution
Lane 1: C6 cell lysate at 10 µg
Lane 2: PC-12 cell lysate at 10 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution
Predicted band size: 34 kDa
All lanes: Western blot - Anti-Cdk2 antibody [E304] (ab32147) at 1/1000 dilution
All lanes: NIH/3T3 cell lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution
Predicted band size: 34 kDa
ab32147 staining Cdk2 in human squamous cell carcinoma of cervix tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/50). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.
All lanes: Western blot - Anti-Cdk2 antibody [E304] (ab32147) at 1/1000 dilution
Lane 1: Jurkat cell lysate at 20 µg
Lane 2: Hela cell lysate at 20 µg
Lane 3: K562 cell lysate at 20 µg
Lane 4: 293 cell lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution
Predicted band size: 34 kDa
All lanes: Western blot - Anti-Cdk2 antibody [E304] (ab32147) at 1/1000 dilution
Lane 1: Human osteosarcoma whole cell lysate - control, non-targeting siRNA at 20 µg
Lane 2: Human osteosarcoma whole cell lysate - siRNA for CDK2 at 20 µg
All lanes: HRP-conjugated goat anti-rabbit IgG polyclonal at 1/2000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 34 kDa
Exposure time: 2s
ab32147 staining Cdk2 in human tonsil tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/50). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.
This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab32147 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
All lanes: Western blot - Anti-Cdk2 antibody [E304] (ab32147) at 1/1000 dilution
Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 10 µg
Lane 2: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 34 kDa
Exposure time: 4min
ab32147 staining Cdk2 in the HeLa cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/200). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 555) ab150078 (1/500) an Alexa Fluor® 555-conjugated Goat anti-rabbit IgG was used as the secondary antibody. Nuclei were counterstained with DAPI.
Western blot: Anti-CDK2 antibody [E304] (ab32147) staining at 1/2000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab32147 was shown to bind specifically to CDK2. A band was observed at 34 kDa in wild-type MCF7 cell lysates with no signal observed at this size in CDK2 knockout cell line Human CDK2 knockout (hetero) MCF7 cell line ab282628. To generate this image, wild-type and CDK2 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Cdk2 antibody [E304] (ab32147)
Lane 1: Wild-type MCF7 cell lysate at 10 µg
Lane 2: CDK2 knockout MCF7 cell lysate at 10 µg
All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling Cdk2 with purified ab32147 at 1/80 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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